Palatable micro-capsules

ABSTRACT

A formulation comprising at least one active or at least one ingredient, and a plurality of micro-capsules formed from a plurality of micro-organisms and having at least one flavouring encapsulated and passively retained within said micro-capsules, said flavouring not being a natural constituent of said micro-organisms, said micro-capsules having: (a) an at least substantially intact cell wall; and (b) an intact cell membrane; wherein said at least one active or said at least one ingredient is foul-tasting and said at least one flavouring masks, disguises or neutralises the foul-taste of said at least one active or said at least one ingredient, preventing a patient to whom said formulation is orally administered from tasting said foul-tasting active, or causing the flavour of said foul-tasting at least one active or ingredient to be reduced such that it is more palatable. Also disclosed are methods of manufacturing said formulations.

This invention relates generally to microbial micro-capsule formulationswhich contain foul tasting and/or foul-smelling ingredients or activeingredients (actives), which have been rendered palatable through themodulation of flavour, odour, texture, colour, temperature and/orviscosity.

Patents FR 2179528, U.S. Pat. No. 4,001,480, EP 0085805, GB 2162147 andEP 0242135, all describe methods/processes for the encapsulation ofsmall molecules including actives inside micro-organisms such as yeastor bacteria.

Oral administration of actives to a patient currently represents themost convenient, cost effective and preferred form of drug delivery. Thehuman tongue contains over 9000 taste buds which distinguish salt,bitter, sour, sweet and umami tastes. It will be appreciated that amajor requirement of an orally administered active that contacts thetaste buds is that the dosage form must be palatable, since anunpalatable formulation increases the risk of a patient neglecting totake the active. Such non-compliance with the dosing regimen can delayor prevent the patient's recovery from the condition under treatment.

Many useful, effective actives have a bitter taste when dissolved inliquid form or even when administered as pills or tablets. For example,a single Ciprofloxacin hydrochloride particle dissolving in the mouthhas such a bitter, unpleasant taste that the product is invariablyrejected by the patient. In the case of those compounds which haveunpleasant (e.g. bitter) taste and/or odour characteristics, theprovision of a dosage form represents a considerable problem. Withimprovements in flavour technology, patients now expect and demandorally administered medications that are pleasantly, or at leasttolerably, flavoured. This is especially true with children and olderadults for whom solutions are most often prescribed. Consequently, thepalatability of orally administered actives is a major concern in thepharmaceutical industry.

Palatability of an orally administered active is influenced by acombination of sensory perceptions including taste and smell, and to alesser extent, texture, appearance, and temperature of the formulation.In the case of microbial micro-capsules, although the micro-organism isusually intact or substantially intact after encapsulation of theactive, in formulations where microcapsules are administered orally andthe encapsulated active is foul-tasting, patients will detect theundesirable and unpleasant taste of the active within the microcapsules.

The present inventors have sought to provide formulations for oraladministration wherein any unpleasant taste and/or odour characteristicsof the active or ingredients are masked, disguised or neutralisedthrough the use of an encapsulated flavouring, or combination offlavourings, which helps ensure quantitative intake of the intendeddose, thereby reducing patient non-compliance.

According to a first aspect of the present invention there is provided aformulation comprising at least one active or at least one ingredient,and a plurality of micro-capsules formed from a plurality ofmicro-organisms and having at least one flavouring encapsulated andpassively retained within said micro-capsules, said flavouring not beinga natural constituent of said micro-organisms, said micro-capsuleshaving:

(a) an at least substantially intact cell wall; and

(b) an intact cell membrane;

wherein said at least one active or said at least one ingredient isfoul-tasting and said at least one flavouring masks, disguises orneutralises the foul-taste of said at least one active or said at leastone ingredient, preventing a patient to whom said formulation is orallyadministered from tasting said foul-tasting active, or causing theflavour of said foul-tasting at least one active or ingredient to bereduced such that it is more palatable.

The term “active” as used herein is meant to include any drug, ortherapeutic or otherwise active agent, preferably a pharmaceuticalcompound or chemical that is capable of being orally administered.Illustrative categories and specific examples of actives useful inconjunction with the present invention include: anti-viral agents,analgesics, anaesthetics, anorexics, anti-arthritics, anti-depressants,anti-diabetic agents, anti-inflammatory agents, anti-helminthics,anti-parkinsonism drugs, anti-pruritics, cardiovascular drugs,anti-hypertensives, ACE inhibitors, hormones, immunosuppressives, musclerelaxants, parasympatholytics, parasympathomimetics, psychostimulants,anti-tuberculosis agents, anti-tussives, such as dextromethorphan,dextromethorphan hydrobromide, noscapine, carbetapentane citrate, andchlophedianol hydrochloride; histamine H1-receptor antagonists, such aschlorpheniramine maleate, phenindamine tarrate, pyrilamine maleate,doxylamine succinate and phenyltoloxamine citrate; histamine H2-receptorantagonists, such as ranitidine, famotidine, cimetidine, nizatidine androxatidine; decongestants, such as phenylephrine hydrochloride,phenylpropanolamine hydrochloride, pseudoephedrine, hydrochlorideephedrine; various alkaloids, such as codeine phosphate, codeinesulphate and morphine; mineral supplements such as potassium chlorideand calcium carbonates, magnesium oxide and other alkali metal andalkaline earth metal salts; laxatives, vitamins; antacids; ion exchangeresins such as cholestyramine; anti-cholesterolemic and anti-lipidicagents such as gemfibrozil; anti-arrhythmics such asN-acetyl-procainamide; anti-pyretics such as acetominophen, aspirin; nonsteroidic anti-inflammatory (NSAI) substances, and more particularlyarylcarboxylic derivatives such as ibuprofen, ketoprofen, flurbiprofen,diclofenac, etodolac and naxoprene; NSAI oxicam derivatives such aspiroxicam, meloxicam, tenoxicam, NSAI fenarnate, indolic, andphenylbutazone derivatives; appetite suppressants such asphenylpropanolamine hydrochloride or caffeine; and expectorants such asguaifenesin. Additional useful active medicaments include coronarydilators, cerebral dilators, peripheral vasodilators, anti-infectives,psychotropics, anti-manics, stimulants, gastrointestinal sedatives andbandages, anti-diarrhoeal and anti-constipation preparations,anti-anginal drugs, vasodilators, anti-hypertensive drugs,vasoconstrictors and migraine treatments, antibiotics, tranquillisers,anti-psychotics, anti-tumour drugs, anti-coagulants, and anti-thromboticdrugs, hypnotics, sedatives, anti-emetics, anti-nauseants,anti-convulsants, neuromuscular drugs, hyper- and hypoglycaemic agents,thyroid and anti-thyroid preparations, diuretics, anti-spasmodics,uterine relaxants, nutritional additives, anti-obesity drugs, anabolicdrugs, erythiropoietic drugs, anti-asthmatics, anti-histaminic oranti-cholinergic or opiate derivatives (such as codeine,dextromethorphan, ethylmorphine, noscapine, pholcodine), coughsuppressants, oral mucolytics (such as acetylcisteine, ambroxol,bromhexine, carbocisteine, erdosteine, letosteine), anti-uricemic drugsand the like. Other examples of actives are well known to a personskilled in the art.

The term “ingredient” is intended to include vitamins, nutritionalsupplements and nutriceutical products.

In this specification, the term “flavouring” generally refers to anysubstance used to improve, enhance, disguise, or mask the taste or odourof an active or ingredient contained within a formulation.

The term “foul-tasting” generally refers to an unpleasant taste and/orodour of an active or ingredient, as perceived by a patient to whom theactive or ingredient is administered. The foul-taste may be due tobitter and/or salty and/or sour characteristics of the active oringredient.

Microbial micro-capsules can be formulated for oral administration to apatient in a variety of ways, for example as a mouthwash, toothpaste,solution, suspension, gel, paste, powder, aerosol, tablet, chewabletablet, capsule, spray, lozenge, syrup, chewing gum, boiled sweet, orcompressed sweet. The use of different formulations are well known to aperson skilled in the art (Remigton's Pharmaceutical Sciences and USPharmacopoeia, 1984, Mack Publishing Company, Easton, Pa., USA; UnitedStates Pharmacopoeia, ISBN: 1889788031). For example, in the case of atablet, the micro-capsules containing encapsulated flavour may be usedto coat the tablet, so that the micro-capsules contact the saliva andmucous membranes of the mouth, rather than the foul-tasting active.

The release of flavouring from microbial micro-capsules (yeast, fungi,bacteria, protozoa, and other unicellular organisms, including microbialderived materials which retain the cell wall structure such as thatdescribed in patent EP 0553176) can occur without physical breakage ofthe cell wall or chemical or biological degradation of the cell wall.Indeed, flavouring is released in a burst of activity when themicro-capsules are placed on a biological membrane such as the membranecoating the tongue. The burst of flavour activity associated with themicro-capsules contacting e.g. the tongue means that the flavour thepatient experiences is an overwhelming taste of the flavouring ratherthan the foul-tasting active or ingredient. The foul-taste of the activeor ingredient is masked, disguised or neutralised, typically completely,by the flavouring, and is not merely diluted by the flavouring. In thecase of yeast micro-capsules for example, the yeast agglomerates (forexample a spray dried agglomerated particle of an average diameter 30microns) may contain a few hundred cells—these agglomerates readilydisperse in the mouth when in contact with saliva down to individualcells and multiples of two or three cells allowing speedy contact andrelease of flavour, overcoming the slower reacting foul-tastingingredient or active.

Preferably the flavouring is lipophilic—i.e. it is soluble within thelipid membrane of the micro-organism used for encapsulation.

Micro-capsules may be formulated wherein the at least one active or atleast one ingredient is encapsulated and passively retained within themicro-capsules, the at least one active or the at least one ingredientnot being a natural constituent of the micro-organisms.

Micro-capsules may be formulated with an at least one additionalflavouring to further mask foul-taste and improve the palatability ofthe formulation. For example, in the case of micro-capsules (containingan encapsulated flavouring such as orange oil) formulated into agelatin, glycerine, or glycerol mono-stearate capsule, the additionalflavouring may be incorporated into the gel matrix of the capsule shell.The additional flavouring may also be e.g. orange oil. The use of anencapsulated flavour combined with an additional flavour is termed ‘dualmasking’, and this technique improves taste and acceptability during andafter swallowing, when a rebound aftertaste may occur. Such ‘dualmasking’ is applicable in a wide range of preparations which containstrong tasting ingredients, such as fish oil, or garlic oil/powder,thereby eliminating the need for a deodorising process.

The flavouring may prevent a patient to whom the formulation is orallyadministered from tasting said foul-tasting ingredient or active, or theflavouring may reduce or lessen the foul taste associated with theingredient or active, making the administered formulation morepalatable.

There is often a correlation between the chemical structure of an activeand its taste. Low molecular weight salts tend to taste salty wherehigher molecular weight salts tend toward bitterness. Nitrogencontaining compounds, such as the alkaloids, tend to be quite bitter.Organic compounds containing hydroxyl groups tend to become increasinglysweet as the number of hydroxyl groups increase. Organic esters,alcohols, and aldehydes are known for their pleasant taste and coolsensation produced by their volatility. The determination of afoul-taste of an active is carried out by standard, well-knownpractices, and is a characteristic often listed along with a descriptionof the active in texts such as The Merck Index, 11th ed., S. Budavari etal. eds., Merck & Co., Inc., Rahway, N.J. (1989) and Rernington'sPharmaceutical Sciences, 18th ed., A. Gennaro ed., Mack Publishing Co.,Easton, Pa. (1990).

Examples of bitter and/or unpleasant tasting actives applicable totaste-masking are: Histamine H₂-antagonists, such as, for example,cimetidine, ranitidine, famotidine, nizatidine, etinidine, lupitidine,nifentidine, niperotidine, roxatidine, sulfotidine, tuvatidine andzaltidine; Antibiotics, such as penicillin, ampicillin, chloramphenicol,erythromycin, ciprofloxacin, norfloxacin, roxithromycin, cloxacillin andclarithromycin; Antimalarials such as chloroquin phosphate and quininesulphate; Decongestants such as pseudoephedrine hydrochloride;Prokinetics such as metoclopramide hydrochloride; Antihistamines such asdiphenhydramine, terfenadine, phenothiazine and chlorpheniramine;Nonsteriodal anti-inflammatory drugs (NSAIDs) such as ibuprofen,acetaminophen, nabumetone and naprosyn; Cough suppressants such asdextromethorphan hydrobromide. Other bitter and unpleasant tasting drugsinclude, caffeine, theophylline (asthma), spironolactone (aldosteroneantagonist), guaifenesin (expectorant), prednisolone (corticosteroid),methacholine (bronchial challenge drug), neostigmine(acetylcholinesterase inhibitor), epinephrine (sympathomimetic),albuterol (Antiasthmatic, broncodilator; antihypertensive),chlorpromazine (sedative), chlordiazepoxide (librium—sedative, hypnotic,anxiolytic and muscle relaxant), amitriptyline (antidepressant),barbiturates, diphenylhydantoin (anticonvulsant), morphine (narcoticanalgesic), meperidine (narcotic analgesic), lomotil (anti-diarrhoea),lidocaine (local anaesthetic). The above actives are not limiting andother foul-tasting actives will be well known to a person skilled in theart.

The flavouring may be one or more flavouring oils. For the purpose ofthis invention, flavouring oils used herein refer to both entireessential oils and the aroma chemicals malting up the essential oils.Essential oils are predominately volatile materials from botanicalsources. The most widely used process for the isolation of essentialoils is steam distillation of plant matter, although dry distillationand solvent extraction are also used. Essential oils are generallyrecognized as safe compositions that can be included in ingestedmaterials. Aroma chemicals refer to chemicals which may be synthetic ornatural, derived from essential oils, i.e., derived from plants bydistillation, expression, or extraction, and which usually carry theflavour of the plant from which they are derived.

Although the invention is not limited to the specific essential oilslisted individually in this specification, a number of importantessential oils include: Almond-Bitter oil, Anise oil, Anise Star Darkoil, Gurjun Balsam oil, White Gurjun Balsam, Basil oil, Bergamot oil,Camphor oil, Caraway oil, Cassia oil, Cananga oil, Chamomile oil, Cherryoil, Cinnamon oil, Citronella oil, Clove Stern oil, Clove Leaf oil,Clove Bud oil, Cognac oil, Coriander oil, Cubeb oil, Eucalyptus oil,Eugenol oil, Ginger oil, Grapefruit oil, Jasmine oil, Laurel oil,Lavender oil, Lemon oil, Lime oil, Mace oil, Mandarin oil, Mayonara oil,Menthol oil, Mint oil, Nutmeg oil, Orange oil, Patchouli oil, PeppermintYakima oil, Peppermint oil, Rose oil, Sage oil, Sassafras oil, Spearmintoil, Tangerine oil, Thyme oil, Violet oil, Vetiver oil, or Wintergreenoil.

Aroma chemicals include but are not limited to anethole, carvone,cintronellal, and camphor.

The flavouring may be one or more natural essences, for example anessence derived from Coffee, Tea, Chamomile, Cocoa, Ginger, Grape,Hazelnut, or Guava.

The flavouring may be one or more natural extracts. For example, theflavouring may be Almond Extract, Anise Extract, Caraway Extract,Cardamom Extract, Celery Seed Extract, Chocolate Extract, CinnamonExtract, Clove Extract, Coriander Extract, Dark Cocoa Extract, GrandMarnier Extract, Lemon Extract, Lemon Lime Extract, Lime Extract,Mandarin Mint Extract, Orange Blossom Extract, Orange Extract, ParsleyHerb Extract, Rum Extract, Tangerine Extract, Tarragon Extract, orVanilla Extract Bourbon.

The flavouring may be one or more artificial flavourings, or naturalflavourings.

For veterinary applications, the flavouring may be a savoury flavourwhich would be appealing to livestock and domestic animals such as dogsand cats. For example, the flavouring may be beef, roast beef, pork,ham, chicken, fish, crab, lobster, shrimp, or scallops.

The flavouring may be a spicy flavour, such as cinnamon, clove, jalapenopepper, mace, or nutmeg.

The flavouring may be a nutty flavour. The flavouring may be almond,butter pecan, cashew, coconut, English walnut-black, hazelnut, peanut,pecan, pistachio, walnut, and walnut-black.

Alternatively, the flavouring may be one or more flavourings widely usedin the pharmaceutical industry. Typically, such pharmaceuticalflavourings have long lasting taste profiles and well characterisedtaste masking properties. Pharmaceutical flavourings include: Anise,Apple, Apricot, Banana, Blackberry, Blueberry, Brandy, Butter, ButterRum, Butterscotch, Caramel, Champagne, Cherry-Black, Cherry-Maraschino,Cherry-Red, Cherry-Wild, Cherry Apricot, Cherry Mint, Coconut, Coffee,Cognac, Cola, Cranberry, Cream Soda, Currant-Black, Egg Nog, Fennel,Ginger Ale, Grape, Grapefruit, Grenadine, Hazelnut, Lemon, Lemon-Lime,Maple, Maple Walnut, Mint Orange, Passion Fruit, Peach, Pineapple, Plum,Prune, Raspberry, Root Beer, Rum, Rum & Coffee, Sherry, Spearmint,Tangerine, Tutti Frutti, or Vanilla Custard.

Flavourings such as aroma chemicals, natural essences, essential oils,natural extracts, artificial flavours, natural flavours andpharmaceutical flavours are commonly available, for example from BluePacific Flavours & Fragrances, Inc., (1354 South Marion Court, City ofIndustry, Calif., USA 91745-2418, www.bluepacificflavors.com).

The flavouring may be a sweetener. Sweeteners can be both natural andsynthetic. In this specification, the term “sweetener” refers to sweetsubstances which are conventionally known or will possibly be known inthe future. Examples of the sweet substances include sucralose,alpha-glucosyltransferase-treated stevia, alpha-cyclodextrin,beta-cyclodextrin, aspartame, acesulfame potassium, N-acetylglucosamine,arabinose, alitame, isotrehalose, isomaltitol, isomaltooligosaccharide(isomaltose, isomaltotriose, panose, etc.), erythritol,oligo-N-acetylglucosamine, galactose, galactosylsucrose,galactosyllactose, galactopyranosyl (beta 1-3) galactopyranosyl (beta14) glucopyranose, galactopyranosyl (beta 1-3) glucopyranose,galactopyranosyl (beta 1-6) galactopyranosyl (beta 1-4) glucopyranose,galactopyranosyl (beta 1-6) glucopyranose, glycyrrhiza extract(glycyrrhizin), xylitol, xylose, xylooligosaccharide (xylotriose,xylobiose, etc.), glycerol, triammonium glycyrrhizinate, tripotassiumglycyrrhizinate, trisodium glycyrrhizinate, diammonium glycyrrhizinate,dipotassium glycyrrhizinate, disodium glycyrrhizinate, curcurin,glucose, gentiooligosaccharide (gentiobiose, gentiotriose,gentiotetraose, etc.), saccharin, sodium saccharin, cyclamate, sucrose,stachyose, stevia extract, powdered stevia, dulcin, sorbitol, sorbose,thaumatin, Theander oligo saccharide, tenryocha extract, trehalulose,trehalose, monellin, nigerooligosaccharide (nigerose, etc.), neotame,neotrehalose, palatinit, palatinose, palatinosepalatinoseoligosaccharide, palatinose syrup, fucose, fructooligosaccharide(kestose, nystose, etc.), fructosyl transferase-treated stevia,fructofuranosyl nystose, Brazilian licorice extract, fructose,polydextrose, maltitol, maltose, maltosyl beta-cyclodextrin,maltotetraitol, maltotriitol, maltooligosaccharide (maltotriose,tetraose, pentaose, hexaose, heptaose, etc.), mannitol, miracle fruitextract, melibiose, rakanka (Momordica grosvenori)) extract, lactitol,lactulose, lactose, raffinose, rhamnose, ribose, isomerized corn syrup,reduced isomaltooligosaccharide, reduced xylo-oligosaccharide, reducedgentiooligosaccharide, reduced malt sugar syrup, glucose syrup,hydrogenated glucose syrup, enzymatically modified licorice, licoricehydrolysates, coupling sugar, soybean oligosaccharide, inverted sugar,glucose syrup, honey and like sweet substances.

The amount of the flavouring to be used in oral formulations ofmicrobial microcapsules in accordance with this invention may be fixedat any desired (e.g. a conventional) level. Typical flavour loading maybe for example 0.5-2% w/w Peppermint oil, 0.5-1% w/w Lemon oil, 0.5%-1%w/w Strawberry flavour (Ungurer), 0.5-2% Aniseed oil. All flavours aretypically encapsulated to the maximum possible level of encapsulationwithin a 4-hour incubation. Actives may be diluted with maltodextrin toa standard 20% W/w flavour loading.

If the encapsulated active is particularly foul tasting (e.g. extremelybitter), the addition of a sweetener such as e.g. aspartame may onlypartially mask or disguise the taste. A further debittering agent, suchas ammonium glycyrrhizinate may be used to more fully mask or disguisethe taste of the active. Ammonium glycyrrhizinate may be used at aweight ratio to the otherwise bitter-tasting active of about 1:50 toabout 1:10, and most preferably at weight ratio of about 1:20 ammoniumglycyrrhizinate to active. Ammonium glycyrrhizinate is the mono-ammoniumsalt of a triterpenoid saponin that consists of an aglycone ofglycyrrhetic acid and a sugar moiety of two glucuronic acid units linkedto each other. This material is about 50 to about 100 times sweeter thansucrose, and is known to be useful in masking bitterness.

The bitterness-masking properties of ammonium glycyrrhizinate mayfurther be enhanced through the use of Polyvinylpyrrolidone (PVP)— thesecompounds appear to potentiate each other to provide the desiredbitterness-masking effect. For example, a particularly bitter-tastingactive such as Ciprofloxacin hydrochloride may be formulated withmicrobial microcapsules containing PVP (5-30% by weight), ammoniumglycyrrhizinate (0.01-0.5% by weight), together with one or moreflavourings. Alternatively, the micro-capsules containing flavouring maybe used to coat a tablet for example, which comprises the foul-tastingactive, together with PVP and ammonium glycyrrhizinate.

Discovering the flavouring best suited to masking an unpleasant taste isoften a very empirical matter. However, general guidelines regarding thetype of flavour best suited to mask a given taste are well known to aperson skilled in the art. For example, salty tastes may be maskedthrough the use of cinnamon, raspberry, orange, maple, and butterscotchflavours, bitter tastes may be masked through the use of cocoa,chocolate-mint, wild cherry, walnut, and raspberry flavours, and sourtastes may be masked through the use of fruit, citrus, and cherryflavours.

The age of the patient and the frequency of dosing will furtherinfluence patient taste preferences. Children generally prefer sweet,fruity, and candy-like tastes while adults prefer less sweet,tart-fruity flavours. Flavouring agents are generally unnecessary andnot recommended for infants under 3-6 months of age. Older adults mayprefer mint or even wine flavoured vehicles. Since liquids which requirefrequent administration, such as antacids, may rapidly become tiresomewith a sweet fruity taste, they may be formulated in mint or tart citrusflavours.

Although altering taste perception by masking unpleasant tastes with aflavouring agent is the major factor in producing palatable formulationscontaining microbially encapsulated flavourings, other factorscontribute as well. Odour is a strong determinant of taste perception,and the aroma or scent of any oral solution should be pleasant andshould correlate with the flavour. Since product temperature can alsoinfluence taste perception, refrigerating liquid products can generallyreduce unpleasant taste. Product texture also plays a role in tasteperception and patient acceptance. Gritty or chalky preparations arepoorly received by most patients (as attested to by the large amounts ofmoney spent by antacid manufacturers to promote their non-chalky,non-gritty products).

The role of colour can also be important in determining patientacceptability for a formulation. Clear, water-like solutions may bepoorly accepted on the basis of perceived inertness or lack of potency.Dark colours, often associated with poisons, such as dark purple, navy,black and brown, may also be rejected. More pleasant, fruity colours aregenerally preferred and should be coordinated with flavours and scents(e.g., yellow-lemon, red-cherry). In formulations where masking of thefoul taste of an ingredient or active is only partially possible, anidiosyncratic selection of flavouring, colouring and odour may distractthe patient sufficiently long enough to allow the formulation to beadministered. For example, a particularly bitter active can be partiallymasked in a blue coloured, strawberry flavoured, mint-scentedformulation. The theory is that while the brain reconciles thecontradictions of a minty, blue strawberry, the foul-tasting ingredientor active has been swallowed.

Texture should also be considered in the preparation of any orallyadministered formulation. Viscosity plays an important role in patientacceptance—the characteristic viscosity of syrups appears to have apositive effect on patient acceptance, whereas less viscous solutionsmay be perceived as “watered down” and more viscous solutions asunpleasant. In the formulation of microbial micro-capsules as asuspension it is important to ensure that the particle size issufficiently small enough so that the product is non-chalky ornon-gritty and that the viscosity is such that the product is fluidenough to flow while being viscous enough to maintain particledispersion.

According to a second aspect of the present invention, there is providedthe use of an at least one flavouring in the manufacture of aformulation, said formulation comprising at least one active or at leastone ingredient, and a plurality of micro-capsules formed from aplurality of micro-organisms, said micro-capsules having:

(a) an at least substantially intact cell wall; and

(b) an intact cell membrane;

said at least one flavouring being encapsulated and passively retainedwithin said micro-capsules, said flavouring not being a naturalconstituent of said microorganisms, wherein said at least one active orsaid at least one ingredient is foul-tasting and said at least oneflavouring masks, disguises or neutralises the foul-taste of said atleast one active or said at least one ingredient, preventing a patientto whom said formulation is orally administered from tasting saidfoul-tasting active, or causing the flavour of said foul-tasting atleast one active or ingredient to be reduced such that it is morepalatable.

According to a third aspect of the present invention, there is provideda method of manufacture of a formulation, said formulation comprising atleast one active or at least one ingredient, and a plurality ofmicro-capsules formed from a plurality of micro-organisms and having atleast one flavouring encapsulated and passively retained within saidmicro-capsules, said micro-capsules having:

(a) an at least substantially intact cell wall; and

(b) an intact cell membrane;

said flavouring not being a natural constituent of said micro-organisms,wherein said at least one active or said at least one ingredient isfoul-tasting and said at least one flavouring masks, disguises orneutralises the foul-taste of said at least one active or said at leastone ingredient, preventing a patient to whom said formulation is orallyadministered from tasting said foul-tasting active, or causing theflavour of said foul-tasting at least one active or ingredient to bereduced such that it is more palatable, comprising the steps of:

(i) contacting said micro-organisms with said at least one flavouring,said at least one flavouring being capable of diffusing into the cellwall of said micro-organism without causing total lysation thereof,whereby said at least one flavouring is absorbed by said micro-organismsby diffusion across the cell wall and is retained passively within saidmicro-organism to produce a plurality of micro-capsules containing saidat least one flavouring,

(ii) formulating said micro-capsules with said at least one active oringredient.

The flavouring may be encapsulated according to any of the methodsdescribed in Patents FR 2179528, U.S. Pat. No. 4,001,480, EP 0085805, GB2162147 and EP 0242135. These encapsulation methods are not limiting andother methods may be known to a person skilled in the art.

The micro-organism is preferably a fungus. Typical fungi are yeasts e.g.Saccharomyces cerevisiae (brewer's yeast and baker's yeast),Kluyveromyces fragilis (dairy yeast), Saccharomyces bayanus (wine yeast)and Candida utilis. Yeasts may be selected from the taxonomic orderEndomycetales. The micro-organism may be a filamentous fungus, e.g.Aspergillus niger. The spore, mycelium and giant cell forms offilamentous fungi may be employed. The micro-organism may be a mold,e.g. Fusarium graminearium. Other micro-organisms which may be employedare bacteria and algae. Any relatively large protozoa also may beutilised—examples of such protozoa include Bacteriodes succinogenes,Etidinium ecaudatum, Entodinium caudatum, Eudipolodinium neglectum,Eudiplodinium maggii, Diplodinium dentatum, and Polyplastronmultivesiculatum.

The flavouring should be in liquid form during the encapsulation processand should be soluble in the cell membrane for encapsulation to takeplace. The flavouring may be a liquid (including oil) in its normalstate, or it may be a solid, in which case it should be dissolved ormicro-dispersed in a solvent which is lipid soluble. Suitable solventsinclude:

(a) primary alcohols within the range C4 to C12, such as nonanol anddecanol (higher alcohols containing a linear chain of more than twelvecarbon atoms are too large for encapsulation);

(b) secondary and tertiary alcohols;

(c) glycols such as diethylene glycol;

(d) esters—any ester where the straight carbon chain is greater than 2and less than or equal to 12, e.g ethyl butyrate, triacetin;

(e) aromatic hydrocarbons such as xylene, and acetophenone,

(f) any aromatic lipophilic oil with no straight chain branch greaterthan 12 carbons.

(g) carboxylic acids between C3 and C12.

Solid flavourings may be encapsulated, however it should be lipophilicto encapsulate successfully and it should either melt below 80° C. or besoluble in one of the above solvents. Prolonged temperatures above 80°C. would damage the cell membrane beyond repair. Ideally for the processthe flavouring should be liquid between 40 and 65° C. since highertemperatures may result in degradation of the flavouring.

Multiple flavourings may be co-encapsulated—e.g. a combination ofessential oils.

In instances where the flavouring is an oil, such as peppermint oil, asolid active, ingredient or additional flavouring can be dissolved inthe oil and co-encapsulated with it. The oil serves as a solvent for theactive, ingredient or additional flavouring, and also imparts flavour tothe resulting micro-capsules. The suitability of flavourings forencapsulation may be found by a simple trial of the method of theinvention.

Methanol, ethanol and isopropanol are very low molecular weight volatilesolvents, which can be used to assist in encapsulation but do notactually encapsulate themselves. If used to encapsulate a material theflavouring must be soluble in e.g. ethanol and when added to e.g. 3 or 4parts water the active must stay in solution. There must always be somewater present to swell the yeast thereby hydrating the membrane, orencapsulation will not take place. The ethanol evaporates during theprocess and the flavouring, which must be at least partially solublewithin the yeast membrane, is encapsulated. Residual ethanol willevaporate during post-encapsulation treatments such as spray drying.

Several criteria must be considered in order to predict whether aflavouring can be encapsulated. Flavourings having a benzene ornaphthalene ring appear to be particularly suitable for encapsulation.Actives with an octanol/water (log P) partition coefficient of between0.5 and 4.0 will encapsulate well. Molecular weight must also beconsidered—flavourings with a molecular mass less than 1000 Daltons cangenerally be encapsulated. Size is also important—since straight chainhydrocarbons greater than C12 generally do not encapsulate, any moleculecontaining a straight chain C12 stretch or greater will probably notencapsulate, nor will a molecule with a rigid structure similar inlength to a C12 chain. Molecules with a greater number of carbons thanC12 can be encapsulated as long as the structure contains benzene rings,e.g. phenolics, or naphthalene rings, etc. Molecules with a smallmolecular diameter are typically encapsulated most efficiently. Volatilemolecules with one to three carbons generally do not encapsulate, e.g.ethane, ethanol, propanol, whereas molecules containing four or morecarbon atoms generally do encapsulate. The optimal range forencapsulation in terms of straight chain carbon atoms lies betweenC4-C12. Beyond these criteria, the suitability of flavourings forencapsulation may be found by a simple trial of the method of theinvention.

The encapsulation treatment may be performed at normal ambienttemperatures but preferably the temperature is elevated, in order toexpedite the encapsulation treatment. A suitable elevated temperaturemay be in the range 35 to 60° C., for instance in the range 40 to 50° C.

The encapsulation treatment preferably comprises mixing themicro-organism with the flavouring in a liquid medium, especially anaqueous medium, to attain good dispersion and contact of themicro-organism with the flavouring.

The encapsulation treatment may be continued until optimum encapsulationhas been achieved. Encapsulation may usually be observed microscopicallyas one or more globules of the flavouring contained within the microbialcell, unless the yeast is grown in a harsh environment (such as highalcohol content), in which case the cell wall can be thickened whichmakes direct visualisation by light microscopy more difficult. In suchinstances, transmission electron microscopy (TEM) may be required. Theencapsulation treatment may take a few hours before the optimum level ofencapsulation is achieved.

After encapsulation, residual low molecular weight solvents such asethanol, methanol and propanol may be removed after the encapsulationprocess through processes such as spray drying. Water may also beremoved by spray- or freeze-drying. Water may also be removed byevaporation by putting the micro-capsule suspension in a dry oven.

A pre-treatment bleaching step may be carried out prior toencapsulation. For example, the treatment may be performed at roomtemperature for up to one hour where the micro-organism is treated witha solution of an alkaline bleach solution comprising 0.2 M sodiumhydroxide/1% w/v hydrogen peroxide, with a pH value of between 9-10.Sodium silicate may be added to the mixture as an anti-foam agent. Theresulting micro-organisms are generally off-white in colour, and thecell wall may be more porous. For example, in the case of bleachedyeast, the cells when dry may absorb between 5-10 times their weight inwater, compared to untreated yeast cells which may absorb between 2-3times their weight in water. This increased capacity of the bleachedyeast to absorb water means that encapsulation is usually performed in agreater volume of liquid, thereby avoiding problems associated withincreased viscosity.

Prior to, or in some cases during, the encapsulation process, themicro-organism may be treated at an elevated temperature and/or with anenzyme and/or with a chemical such as a magnesium salt to improve theefficiency of encapsulation. Enzymes such as pepsin, trypsin,chymotrypsin, chitinase, and β-glucanase serve to degrade the microbialcell wall. Magnesium salts enhance permeability of the micro-organism.The micro-organism may then be mixed with the flavouring to beencapsulated and incubated until optimum encapsulation is achieved (asdetermined by light or electron microscopic analysis of themicro-capsules).

After encapsulation of the flavouring, and prior to preparation of theformulation, a conditioning treatment of the resulting micro-capsulesmay be performed to remove any residual microbial taste, colour andodour of the micro-capsules. This conditioning treatment may compriseincubating the micro-capsules in a dry environment such as an oven orheat chamber at room temperature for several weeks or months, or at anelevated temperature of up to 40° C. for hours/days.

In the case of yeast, the encapsulation process results in theaccumulation of actives within the naturally double walled capsule.Yeast cell walls are generally 80-90% polysaccharide, includingpredominant glucans such as 1,3-β-glucan, and also the long chaincarbohydrate polymer chitin which adds rigidity and structural supportto the cells. Proteins (such as mannoproteins), lipids andpolyphosphates together with inorganic ions make up the cell wallcementing matrix. The inner membrane is a typical lipid bilayer. Theyeast cell wall, unlike many food grade capsules, is insoluble andtherefore the micro-capsules can be wet and dry processed. When theyeast microcapsules are spray dried a free flowing powder is producedmade up of agglomerated particles comprising numerous yeast cells.Depending on drying conditions the dry particle size can range between10 and 300 microns. For large particles a fluidised bed is required. Theproduct can also be prepared as a cake, suspension, produced bypressing, or rotary drying. Particle size or a mixture of particle sizesmay be useful to control release rates.

The micro-capsules may be washed after encapsulation to remove residualunencapsulated material and isolated by centrifuging, freeze-drying orspray-drying.

The invention will be further apparent from the following experimentswhich show, by way of example only, formulations of the presentinvention and methods of manufacture of same.

EXPERIMENTS

The following examples detail various formulations of actives andflavourings for oral administration, and the methods used to manufacturesuch formulations.

Example 1

Saccharomyces cerevisiae 62F was sourced from Williams Bio-Energy, PekinIll. The spray dried yeast was pre-washed with water to remove residualmedia components and the resulting washed yeast suspended in water to afinal concentration of 33% w/v. Peppermint oil was added to a finalconcentration equivalent to 50% of the dry weight of the yeast. Themixture was stirred at 45° C. for 6 hours in a water-jacketed vesselusing a Teflon coated paddle. The yeast cells containing encapsulatedpeppermint oil were harvested by centrifugation, 2000×g, and washed withwater twice to remove residual unencapsulated flavour. The washed yeastmicro-capsules containing peppermint oil were diluted to 35% dry solidswith water and spray dried. The resulting powder had an average particlesize of 30 microns, each containing approximately 150-250 yeast cellseach containing peppermint oil. The yeast encapsulated peppermint oilwas found to enhance the perception in the mouth of the peppermint oilcompared to unencapsulated free peppermint oil, by a factor of between 4and 5. When flavour oil at a commonly used level in the pharmaceuticalindustry was added to Chloroquin sulphate, a bitter tastinganti-malarial compound, the Chloroquin sulphate could be easily detectedwhen placed on the tongue. When an equivalent weight ofyeast-encapsulated flavour was used the Chloroquin sulphate was nolonger perceived.

Example 2

Torula yeast (Candida utilis) obtained from Overseal Ltd., was washedwith water to remove free dissolvable nutrients and extraneous material.The resulting washed yeast were adjusted with water to a finalconcentration of 30% dry solids and stirred using a rotary stirrer at380 rpm. To this suspension, Ungurer strawberry flavour (a liquidpreparation) was added to approximately 50% the dry weight of the dryweight of the yeast cells. With continuous stirring the mixture wasincubated in a water jacketed, closed vessel for 3 hours at 55° C. Afterthis time the yeast cells were harvested by centrifugation and freezedried. When placed in the mouth the dry powder (20 mg) produced anintense strawberry flavour, 3-4 times that found with the equivalentfree flavour. The flavour was able to mask Pseudoephedrine HCl, a bittertasting decongestant compound mixed into the powder at up to 15% byweight.

Example 3

Spray dried (400 g) brewers yeast (Saccharomyces cerevisiae NCYC 1335)was suspended in a mixture of 500 ml of saturated acesulfame K and 500ml of saturated sucralose. The mixture was incubated at 30° C. for 1hour and dried by freeze-drying. The resultant product was gently milledto a particle size of 100 microns. The product delivered an intensesweetening effect which was able to overcome the flavour ofMetoclopramide, a bitter tasting prokinetic compound. Microscopicexamination confirmed that not all the sweetener was encapsulated and aportion was located on the surface of the capsules.

The same process was carried out using Williams Bioenergy yeast 62F,which had been bleached using the method described in Example 7. In thiscase 1000 ml of acesulfame K and 1000 ml of sucralose was used becauseof the ability of the bleached yeast to take up greater volumes ofwater. The cell wall is also opened up more effectively allowing morespace to be filled with the sucralose. As before on drying the productcontained a mixture of encapsulated sweetener and unencapsulatedsweetener.

Example 4

As perception of flavour, particularly with children is often due toexpectation based on the colour of the product, during encapsulation oflime oil in yeast, a food grade green dye was added to the mixture at0.1% w/v. In this case 250 ml Lime oil (R. C Treate) was added to 1litre of water containing 500 g of spray dried Williams Bio-Energy yeast62F. The mixture was stirred for 4 hours at 50° C. When washed withWater and dried using spray drying the product contained 24% lime oiland was green in colour.

Example 5

With domestic animals, (e.g. dogs and cats), although the flavours maybe similar the format of delivery may be different. To make foul-tastingactives such as the anti-worming agent dichlorophen more palatable,yeast encapsulated beef flavour was added to the foul-tasting actives.In this case, a yeast was chosen which had an inherently beefy flavour,a distiller yeast sourced from Quest International Ltd. The yeast wasdispersed in water to a final concentration of 32% w/v and the liquidflavour added to 50% the dry weight of the yeast. The mixture wasincubated whilst stirring for 8 hours at 40° C. The yeast cellscontaining the beef flavour were harvested by centrifugation, washedwith water and dried by spray drying. The yeast capsules containedapproximately 15% flavour oil. For dogs, in which the product would notremain in the mouth for a long period and little chewing will takeplace, a thin coating of the yeast encapsulated intensely beefyflavoured product over a tablet of the foul tasting material waseffective. For cats where more chewing behaviour takes place beforeingestion, the intense yeast encapsulated beefy flavour was mixed withno greater than 20% of the foul tasting compound, again which waseffective.

Example 6

Saccharomyces cerevisiae NCYC 1603 was maintained on MYGP agar slopes(0.3% (w/v) each of malt extract and yeast extract, 0.5% bacterialpeptone, 2% (w/v) glucose; 2% (w/v) agar). A loop of yeast wastransferred aseptically to 10 ml MYGP broth, media as above but withoutagar and incubated overnight at 30° C. The broth was asepticallytransferred to a fermenter containing 5-litres working volume of MYGPbroth. The fermenter was incubated for 3 days at 30° C. and the yeastharvested by centrifugation using a MSE Mistral 3000i centrifuge(2000×g). The harvested yeast was washed with water to remove excessmedia and suspended in water to a final solids content of 33% w/v in ajacketed glass vessel, temperature 42° C. The yeast was agitated withtop stirring, Stuart Scientific SS10, with Teflon paddle, atapproximately 300 rpm. Peppermint oil (125 g) and spearmint oil (125 g)were added and the suspension continually stirred for 6 hours. The cellswere harvested by centrifugation, 2000×g using a Mistral 3i Centrifugefor 20 minutes. The pelleted yeast cells containing encapsulatedpeppermint and spearmint oil was washed twice with water and the cellsre-suspended in water to approximately 35% solids. The material was thenspray-dried using a Niro Mobile Minor spray dryer. The peppermint oiland spearmint oil concentration was found to be 15% and 17%,respectively.

The dry cells were coated onto the surface of a preformed tabletcontaining Dextromorphan, a bitter tasting cough suppressant compound.When the surface layer came in contact with the mouth and saliva incombination, an intense flavour was released, greatly facilitatingspeedy swallowing of the tablet, with no bitter taste detected. Aportion of the yeast capsules on the surface coating freely dispersedwithin the mouth, enhancing the taste-masking effect. The high intensityflavour release encouraged saliva production, easing swallowing of thetablet containing the active.

Example 7

Commercially available dry bakers yeast (300 g) (Saccharomycescerevisiae) sourced from DCL Ltd., was suspended in one litre of a 0.2 Msolution of sodium hydroxide in water containing 40 g per litre ofsodium silicate. Hydrogen peroxide was added until the concentrationreached 1% w/v and the resulting suspension was then gently stirred forone hour at room temperature. The yeast was then removed bycentrifugation, washed with water to remove excess bleaching agent anddried by spray drying. The yeast produced was white to off-white incolour and in suspension had a creamy texture. Any yeasty odour had beenremoved. The spray-dried material was stored dry at room temperatureready for future encapsulation processes.

A portion of the suspension before drying, was adjusted to 20% solidswith water. The viscosity of the bleached and deodorised yeast was toogreat to obtain the desired emulsion characteristics using a similarconcentration as the unbleached yeast. The bleached and deodorised yeastsuspension was stirred using a rotary stirrer at 350 rpm for 5 hours at40° C. in the presence of liquid caramel flavour to approximately 50% ofthe weight of dry yeast. The yeast cells containing liquid caramelflavour oil were then removed by centrifugation, washed with water anddried by spray-drying. The dry product contained 26% liquid caramel oil.

The dry capsules were mixed with a crystalline formulation of a bittertasting active (Chloropheniramine maleate—an anti-histaminic), ensuringthat the flavour was in excess of the bitter tasting active. For bestresults the yeast capsules containing flavour were in a ratio of 3:1 orgreater with the bitter tasting active. The mixture of yeastencapsulated flavour and bitter tasting active were formed into a tabletusing standard methods known in the pharmaceutical industry.

Example 8

Saccharomyces bayanus (a wine yeast) was suspended in water to a finaldry-solids concentration of 35% w/v. Caffeine was dissolved inpeppermint oil, to 10% w/v, and mixed with the yeast whilst stirring at350 rpm, to a final concentration of 50% of that of the dry yeast for 8hours at 42° C. With this application care must be taken not use asolution of bitter tasting active of greater than 30% solubility in thepeppermint or other flavour oil carrier. If the concentration of bittertasting compound is greater there is a danger that the bitter tastewould also be enhanced over the ability of the flavour oil to mask thefoul tasting material. The yeast cells containing encapsulatedpeppermint oil/caffeine were harvested by centrifugation, and the cellswere washed twice with water. The cells contained 28% w/w peppermint oiland 3.2% caffeine. After formulating the yeast containing peppermint oiland caffeine into a tablet, the product was rendered palatable.

Example 9

Kluyveromyces lactis, (a commercially available yeast) cultured on wheywas suspended in water to a final dry solids concentration of 33% w/v.The yeast suspension was stirred using a rotary stirrer at 350 rpm for 3hours at 60° C. in the presence of menthol, which had been pre-melted,added to approximately 50% of the weight of dry yeast. The yeast cellscontaining menthol were then removed by centrifugation, washed with warmwater and dried by spray-drying. The dry product contained 28% mentholcrystals. When placed on the tongue there was a intense cooling/mentholflavour detected. Bitter tasting compounds e.g. Ciprofloxacin HCl (anantibacterial) could be added to the mixture to a level of 20% beforeovercoming the intense cooling/menthol flavour.

Example 10

Saccharomyces cerevisiae NCYC 1338, (a non-flocculant yeast) wassuspended in water to a final concentration of 35% dry solids. To thissuspension, five times concentrated lime oil was added to 50% of the dryweight of yeast, and the mixture was agitated using a rotary stirrer at300 rpm. The mixture was stirred for 2 hours at 60° C. in the presenceof a nitrogen sparge, to reduce potential for oxidation. The cells wereharvested by centrifugation at 2000×g and the residual lime oil wasremoved by washing with water. An aliquot of the concentrated pellet wasre-suspended in a saturated solution (approx 30% w/v) of acesulfame-K (asweetener) and incubated with gentle stirring (100 rpm) in a rotarystirrer for 30 minutes until the sweetener had diffused into the yeastcells. The preparation was then dried by spray drying, resulting in apowder with yeast cells containing 28% w/w lime oil and 300 g ofacesulfame-K. On microscopic examination it was noted that not all thesweetener had entered the cell, so the powder included yeast capsulescontaining lime oil in the cell cytoplasm, sweetener in the cell wallmatrix and free sweetener.

The second aliquot of the suspension yeast micro-capsules containinglime oil was dried by spray drying and re-suspended in a saturatedsolution of acesulfame-K. After incubation and drying the relativeconcentration of material entrapped within the yeast cell wall matrixwas greater than when the yeast suspension was mixed with the sweetenersolution.

Both of these products, when formulated with a bitter tasting active(Norflaxacin—an antibacterial), delivered intense flavour and strongsweetening effects thereby masking the foul-taste of the active.

Example 11

Where the yeast used in the process would require to be registered as‘Organic’, for example in nutriceutical applications, Bioreal yeast(Agrano GmbH) can be used. A sample of Bioreal yeast was suspended inwater to a final dry solids concentration of 35% and essential orangeoil was added to a final concentration equal to 45% of the dry weight ofthe yeast. The mixture was incubated whilst stirring for 6 hours at 48°C. After washing and spray-drying, the yeast cells contained 26% orangeoil. The encapsulated flavour produced an intense orange flavour, 4-5times that of the equivalent free oil. The yeast encapsulated orange oilwas able to mask up to 20% addition of Cloxacillin sodium—a bittertasting antibacterial compound.

Example 12

Saccharomyces cerevisiae (62F) was obtained from William Bioenergy as aspray dried powder, this yeast was light in colour and had little yeastflavour due to the chosen culture media, which was based on corn syrup.The dry powder washed with water to remove excess media components andthe resultant yeast, approximately 65% of the dry weight of the spraydried powder, was suspended in water to a final solids content of 35%w/v in a jacketed glass vessel, temperature 42° C. The yeast wasagitated with top stirring, Stuart Scientific SS10, with Teflon paddle,at approximately 300 rpm. Ibuprofen dissolved in peppermint oil (10%w/v) was added to the mixture to approximately half the dry weight ofthe washed yeast and the mixture stirred continuously for 6 hours. Theyeast cells containing peppermint and ibuprofen were then removed bycentrifugation, washed with warm water and dried by spray-drying. Theresulting yeast capsules contained 36% w/w peppermint oil and 3.7% w/wibuprofen. The resulting powder was rendered palatable and the bittertaste of ibuprofen was not detectable.

1. A formulation comprising at least one active or at least oneingredient, and a plurality of micro-capsules formed from a plurality ofmicro-organisms and having at least one flavouring encapsulated andpassively retained within said micro-capsules, said flavouring not beinga natural constituent of said micro-organisms, said micro-capsuleshaving: (a) an at least substantially intact cell wall; and (b) anintact cell membrane; wherein said at least one active or said at leastone ingredient is foul-tasting and said at least one flavouring masks,disguises or neutralises the foul-taste of said at least one active orsaid at least one ingredient, preventing a patient to whom saidformulation is orally administered from tasting said foul-tastingactive, or causing the flavour of said foultasting at least one activeor ingredient to be reduced such that it is more palatable.
 2. Aformulation according to claim 1, said at least one active or said atleast one ingredient being encapsulated and passively retained withinsaid micro-capsules, said at least one active or said at least oneingredient not being a natural constituent of said microorganisms.
 3. Aformulation according to claim 1, wherein said formulation additionallycomprises an at least one additional flavouring.
 4. A formulationaccording to claim 1, in which said at least one flavouring or said atleast one additional flavouring is selected from the group consistingof: essential oils, aroma chemicals, natural essences, natural extracts,sweeteners, artificial flavourings, natural flavourings, pharmaceuticalflavourings, or nature identical flavourings.
 5. A formulation accordingto claim 1, wherein said at least one flavouring or said at least oneadditional flavouring additionally comprises ammonium glycyrrhizinateand/or polyvinylpyrrolidone.
 6. A formulation according to claim 1,selected from the group consisting of: mouthwash, toothpaste, solution,suspension, gel, paste, powder, aerosol, tablet, chewable tablet,capsules, spray, lozenge, linctus, syrup, chewing gum, boiled sweet, orcompressed sweet.
 7. The use of an at least one flavouring in themanufacture of a formulation, said formulation comprising an at leastone active or at least one ingredient, and a plurality of micro-capsulesformed from a plurality of micro-organisms, said micro-capsules having:(a) an at least substantially intact cell wall; and (b) an intact cellmembrane; said at least one flavouring being encapsulated and passivelyretained within said microcapsules, said flavouring not being a naturalconstituent of said micro-organisms, wherein said at least one active orsaid at least one ingredient is foul-tasting and said at least oneflavouring masks, disguises or neutralises the foul-taste of said atleast one active or said at least one ingredient, preventing a patientto whom said formulation is orally administered from tasting saidfoul-tasting active, or causing the flavour of said foultasting at leastone active or ingredient to be reduced such that it is more palatable.8. A method of manufacture of a formulation, said formulation comprisingat least one active or at least one ingredient, and a plurality ofmicro-capsules formed from a plurality of micro-organisms and having atleast one flavouring encapsulated and passively retained within saidmicro-capsules, said micro-capsules having: (a) an at leastsubstantially intact cell wall; and (b) an intact cell membrane; saidflavouring not being a natural constituent of said micro-organisms,wherein said at least one active or said at least one ingredient isfoul-tasting and said at least one flavouring masks, disguises orneutralises the foul-taste of said at least one active or said at leastone ingredient, preventing a patient to whom said formulation is orallyadministered from tasting said foul-tasting active, or causing theflavour of said foul-tasting at least one active or ingredient to bereduced such that it is more palatable, comprising the steps of: (i)contacting said micro-organisms with said at least one flavouring, saidat least one flavouring being capable of diffusing into the cell wall ofsaid micro-organism without causing total lysation thereof, whereby saidat least one flavouring is absorbed by said micro-organisms by diffusionacross the cell wall and is retained passively within saidmicro-organism to produce a plurality of micro-capsules containing saidat least one flavouring, (ii) formulating said micro-capsules with saidat least one active or ingredient.
 9. A method of manufacture of aformulation according to claim 8, wherein said micro-organisms areadditionally contacted with an at least one active or an at least oneingredient, wherein said at least one active or said at least oneingredient is foul-tasting, and is not a natural constituent of saidmicro-organisms, said at least one active or said at least oneingredient being encapsulated and passively retained within saidmicro-capsules.
 10. A method of manufacture of a formulation accordingto claim 8, wherein said formulation additionally comprises an at leastone additional flavouring.
 11. A method of manufacture of a formulationaccording to of claim 8 wherein said at least one flavouring or said atleast one additional flavouring additionally comprises ammoniumglycyrrhizinate and/or polyvinylpyrrolidone.
 12. A formulation accordingto claim 1, wherein said micro-organism is selected from the groupconsisting of-: fungus, bacterium, alga and protozoa.
 13. A formulationaccording to claim 1, wherein said micro-organism is a yeast selectedfrom the taxonomic order Endomycetales.
 14. A method of manufactureaccording to claim 6, wherein said micro-organism is selected from thegroup consisting of: fungus, bacterium, alga and protozoa.
 15. A methodof manufacture according to claim 6, wherein said micro-organism is ayeast selected from the taxonomic order Endomycetales.